Use of a stackable surgical guide to improve the accuracy of the lateral wall approach for sinus grafting in the presence of a sinus septum.

The identification and management of interfering maxillary sinus septa is essential to anticipate and prevent membrane perforation and other complications during sinus grafting. A computer-guided sinus approach based on a new magnetic stackable surgical guide was planned, to transfer the exact position of the septum and optimize the positioning of the lateral access windows. This technique reduces the risk of sinus membrane injury, thereby increasing the safety and efficacy of the procedure.

Overcoming the ceiling effects of experts' motor expertise through active haptic training.

One of the most challenging issues among experts is how to improve motor skills that have already been highly trained. Recent studies have proposed importance of both genetic predisposition and accumulated amount of practice for standing at the top of fields of sports and performing arts. In contrast to the two factors, what is unexplored is how one practices impacts on experts' expertise. Here, we show that training of active somatosensory function (active haptic training) enhances precise force control in the keystrokes and somatosensory functions specifically of expert pianists, but not of untrained individuals. By contrast, training that merely repeats the task with provision of error feedback, which is a typical training method, failed to improve the force control in the experts, but not in the untrained. These findings provide evidence that the limit of highly trained motor skills could be overcome by optimizing training methods.

Pacemaker-associated infection caused by ST81/SCCmec IV methicillin-resistant, vancomycin-intermediate Staphylococcus aureus in Japan.

A 76-year-old Japanese man was admitted to hospital for treatment of fever and skin lesion at the implantation site of his pacemaker. During his hospitalization, vancomycin-intermediate Staphylococcus aureus (MIC 4 µg/mL) with reduced susceptibility to daptomycin was isolated from venous blood. This isolate was identified as methicillin-resistant S. aureus with SCCmec IV and was genotyped as sequence type 81, coa VIIa and spa type t7044, harbouring blaZ, aac(6')-aph(2″) and enterotoxin(-like) genes sea, seb, sek, sel, selx and selw. The patient was successfully treated with daptomycin, minocycline and sulfamethoxazole/trimethoprim. We describe the identification of sequence type 81/SCCmec IV vancomycin-intermediate S. aureus from pacemaker-associated septicaemia.

Isolation of dermatan sulfate with high heparin cofactor II-mediated thrombin-inhibitory activity from porcine spleen.

We prepared dermatan sulfate specimens from various porcine tissues, and compared their heparin cofactor II-mediated thrombin-inhibitory activities and chemical natures, including disaccharide composition. Electrophoresis of the specimens on cellulose acetate membrane indicated that spleen dermatan sulfate was the most acidic of the dermatan sulfates prepared from the various porcine tissues. Analysis of the disaccharide units of the dermatan sulfate specimens by high-performance liquid chromatography revealed that spleen dermatan sulfate was rich in 4,6-di-O-sulfated N-acetylgalactosamine residues as compared with those of the other tissues. Spleen dermatan sulfate exhibited the highest thrombin-inhibitory activity, which may be related to its high content of the disulfated N-acetylgalactosamine residue.

The Tokyo subway sarin attack--lessons learned.

The sarin gas attack in the Tokyo subway system is reviewed from a clinical toxicology perspective. Based on the lessons learned from this attack, the following areas should be addressed on a global scale. First, an adequate supply of protective equipment is required, including level B protective equipment with a pressure demand breathing apparatus. In addition, a system should be established that enables a possible cause to be determined based on symptoms, physical findings, general laboratory tests, and a simple qualitative analysis for poisonous substances. If an antidote is needed, the system should enable it to be administered to the victims as quickly as possible. Preparation for a large-scale chemical attack by terrorists requires the prior establishment of a detailed decontamination plan that utilizes not only mass decontamination facilities but also public facilities in the area. A system should be established for summarizing, evaluating, and disseminating information on poisonous substances. Finally, a large-scale scientific investigation of the Tokyo sarin attack should be conducted to examine its long-term and subclinical effects and the effects of exposure to asymptomatic low levels of sarin.

Relation between glucose-stimulated insulin secretion and intracellular calcium accumulation studied with a superfusion system of a glucose-responsive pancreatic beta-cell line MIN6.

The concept that cytosolic free calcium is the primary signal for insulin secretion is generally accepted, but studies with intact pancreatic beta-cells of the cytosolic free calcium concentration-insulin secretion relationship have produced contradictory and sometimes confusing data. We designed a superfusion system of a pancreatic beta-cell line, MIN6, loaded with fura-2, which allowed simultaneous measurement of cytosolic free calcium concentration and insulin secretion. MIN6 cells released insulin in response to high glucose, thus resembling events in normal islet cells. Cytosolic free calcium concentration and insulin secretion rapidly increased, and the increase was suppressed by mannoheptulose or by sodium azide. This increase was suppressed by lowering the temperature of the medium. Cytosolic free calcium concentration and the insulin secretion induced by leucine were not influenced by mannoheptulose but were inhibited by sodium azide. In RINm5F cells, cytosolic free calcium concentration and insulin release were slightly suppressed by glucose but were increased by ionomycin. There was a close relation between the rise in cytosolic free calcium concentration and insulin secretion in all cases. Our findings provide a direct evidence that a rise in cytosolic free calcium concentration depends on glucose metabolism and is a primary signal for insulin secretion.

Biphasic insulin response to high glucose and a role of protons and calcium.

In order to determine the role of protons and Ca++ in the biphasic insulin response to glucose, we studied the effect of monensin, a carboxylic ionophore, on the first phase and second phase of glucose-induced insulin release and Ca++ efflux from perifused rat pancreatic islets. The agent, 1-100 nM, dose dependently inhibited the insulin release from the islets incubated for 60 min in Krebs-Ringer bicarbonate buffer containing 16.7 mM glucose. Islet 14CO2 production rates from D-(U-14C)glucose were not affected by 10 or 100 nM monensin. Perifusion of the islets prelabeled with 45Ca++ demonstrated that 100 nM monensin had only a slight inhibitory effect on the first phase insulin response to 16.7 mM glucose and no effect on 45Ca++ efflux. This agent inhibited the second phase insulin release and depressed 45Ca++ efflux. When monensin was added at the start of the second phase release, the release was inhibited. When exposed to the agent before the stimulation by glucose, the first phase insulin release was observed, albeit significantly decreased, and the start of the insulin release and 45Ca++ efflux was delayed. The agent, added 30 min after the change to high glucose, immediately inhibited the insulin release. Thus, the first phase insulin release is mediated mainly through a mechanism which is not related to protons generated from glucose metabolism. Protons may be a crucial coupling factor in the second phase insulin release.

Intraperitoneal PACAP administration decreases blood glucose in GK rats, and in normal and high fat diet mice.

PACAP is an islet peptide that serves as an endogenous amplifier of glucose induced insulin secretion. Furthermore, we has recently found that PACAP also potentiates insulin stimulated glucose uptake in adipocytes. Therefore, an antidiabetic action of PACAP is possible. In the present study, we examined the effect of PACAP treatment of the hyperglycemia in GK rats, an animal model of type 2 diabetes, and in high fat fed C47BL/6J mice, an animal model for glucose intolerance. GK rats housed with normal diet exhibited a normal level of blood glucose until three weeks old but significant hyperglycemia at eight weeks. When GK rats were treated with daily PACAP38 (i.p. injection, 6 pmol/kg) from age three weeks, development of hyperglycemia was prevented. In high fat fed mice, i.p. administration of PACAP27 for five (25 nmol/kg twice daily) reduced plasma glucose levels to 6.9 +/- 0.2 mmol/l compared to 8.1 +/- 0.2 mmol/l in saline injected animals (p < 0.001) without altering baseline insulin levels. We conclude that PACAP reduces circulating glucose in animal models of type 2 diabetes and glucose intolerance. The mechanism of this action needs to be established.

Current status of PACAP as a regulator of insulin secretion in pancreatic islets.

PACAP-27 and PACAP-38 as low as 10(-13) M stimulate insulin release from rat islets in a glucose-dependent manner. PACAP also glucose dependently increases cAMP and [Ca2+]i in rat islet beta cells. The [Ca2+]i and insulin secretory responses to PACAP exhibit a similar concentration-response relationship, exhibiting a peak at 10(-13) M. When the [Ca2+]i response is abolished by nitrendipine, a blocker of L-type Ca2+ channels, the insulin response is also inhibited. Insulinotropic peptides glucagon, GLP-1, and VIP also increase [Ca2+]i in beta cells, but only in the nanomolar concentration range. PACAP is 4 logs more potent that VIP, a peptide that exhibits 68% amino acid homology and shares the type II PACAP receptor with PACAP. Immunoreactivity for the type I PACAP receptor is demonstrated in rat islets. Furthermore, PACAP immunoreactivity is demonstrated in nerve fibers and islets in rat pancreas. Based on these findings, we can draw the following conclusions: (1) PACAP is localized in pancreatic nerve fibers and islets; (2) PACAP in the subpicomolar range stimulates insulin release from islets; (3) the stimulation of insulin release is mediated by the cAMP-dependent increase in [Ca2+]i in beta cells; (4) all the PACAP effects are glucose-dependent; (5) PACAP is the most potent insulinotropic hormone known, and (6) the type I PACAP receptor appears to mediate the action of PACAP in the subpicomolar range. Finally, we hypothesize that PACAP is a pancreatic peptide of both neural and islet origin and functions as an intrinsic potentiator of glucose-induced insulin secretion in pancreatic islets (FIG 6).

Abnormal calcium handling by perifused pancreatic islets from neonatal streptozotocin diabetic model rats.

To elucidate the mechanism of impaired insulin release in case of non-insulin-dependent diabetes (NIDDM), we investigated insulin release and 45Ca++ efflux from perifused islets obtained from neonatal streptozotocin diabetic model rats. The model rats were prepared by the intraperitoneal administration of 65 mg/kg streptozotocin (STZ) to neonatal males. Rats treated with STZ did not differ from controls in body weight from 1 week to 16 weeks. The model rats had significant hyperglycemia both in the fasting state and after intraperitoneal administration of 2 g/kg glucose. Although the diameter of the islets from the model rats was not significantly different from that of controls, immunoreactivity to anti-insulin was slightly diminished, and degranulation was slightly observed in B-cells. Insulin content was reduced to 45.6% of the control. Insulin release from the perifused islets of STZ-treated rats responded little to 16.7 mmol/L glucose, but normally to 20 mmol/L arginine in the presence of 5.5 mmol/L glucose. In experiments to test the 45Ca++ efflux from the perifused islets prelabeled with 45Ca++, a rise of 45Ca++ efflux concomitant with the second phase of insulin release from the islets of the model rats was inhibited although a sharp increase of 45Ca++ efflux concomitant with the first phase of insulin release was maintained. 45Ca++ uptake for 30 minutes was reduced in the islets from the model rats in the basal and stimulated state of insulin secretion although the incremental 45Ca++ uptake was similar. It is possible that the abnormal calcium handling in pancreatic B-cells may be one of the causes of defect in insulin release in our model rats.(ABSTRACT TRUNCATED AT 250 WORDS)

First-phase insulin response to glucose in nonobese or obese subjects with glucose intolerance: analysis by C-peptide secretion rate.

This study was proposed to clarify the impairment of first-phase insulin response to glucose in subjects with glucose intolerance by analysis of C-peptide secretion rate after glucose or glucagon injection. The rate was calculated from kinetic analysis of peripheral C-peptide behavior. The rate reached the peak two minutes after glucose injection and then rapidly declined (first-phase secretion) in control subjects. In nonobese subjects with impaired glucose tolerance (IGT) or non-insulin-dependent diabetes mellitus (NIDDM), the rate promptly increased in response to glucose and was followed by a second phase increase. The time course of the rate in the subjects was slightly different from that in control subjects. There was a progressively greater deficit in the first-phase increase with increasing severity of glucose intolerance. The time course of the rate in the obese subjects with NIDDM was different from that in control subjects. The first-phase increase was reduced in the obese subjects with NIDDM. The glucose disappearance rate was correlated with the first-phase increase. Since the time course of the rate after glucagon injection in all subjects did correspond well with that in the control subjects, variation of metabolic clearance rate of endogenous C-peptide among the subjects may be negligible for this study. This study provides the precise time course of first- and second-phase insulin response to glucose injection in nonobese and obese subjects with IGT or NIDDM as well as convincing evidence of the progressive reduction of first-phase insulin response with increasing severity of glucose intolerance. First-phase insulin response to glucose might be slightly delayed in some obese subjects with NIDDM.

Simple method for cryopreservation of an anaerobic rumen fungus using ethylene glycol and rumen fluid.

The cryopreservation of an anaerobic rumen fungus, Piromyces communis OTS1, was examined at -84 degrees C using dimethyl sulfoxide, propylene glycol or ethylene glycol as cryoprotectants. Ethylene glycol was the most effective agent, combining high survival and low toxicity, followed by dimethyl sulfoxide and propylene glycol. Cell-free rumen fluid in the cryopreservation medium decreased the toxicity of the cryoprotectant agents and also had a protective action per se. A survival of 80% after 1 year storage was obtained when samples with an initial zoospore density of 5 x 10(4) zoospores/ml were equilibrated for 15 min in medium containing 0.64 M ethylene glycol and 5% cell-free rumen fluid, then frozen with dry ice and stored at -84 degrees C.

Purification and characteristics of cytosolic chitinase from Piromyces communis OTS1.

A chitinase was purified from the cytosolic fraction of the anaerobic rumen fungus Piromyces communis OTS1 by affinity chromatography using regenerated chitin, gel filtration and chromatofocusing. The chitinase was most active at pH 6.2 and at 60 degrees C in a 20-min assay. The molecular mass of the purified protein was estimated by SDS-PAGE to be 42 kDa and its pI was 4.9. The enzyme activity, which was of the 'endo' type, was inhibited by Ag+, Hg2+ and allosamidin. N-Acetyl-beta-glucosaminidase and 'exo' type chitinase activity were absent from the purified preparation.

Exploratory eye movements and neuropsychological tests in schizophrenic patients.

Exploratory eye movements in schizophrenic and nonschizophrenic subjects were examined with an eye mark recorder while the subjects viewed geometric figures. Elementary components of eye movements and the responsive search score (RSS), a function of the number of sections on which the subjects fixated, were measured by means of an eye movement analyzer and slow motion replay. The schizophrenic group and the depressed patient group had fewer eye fixations than the normal control group and the obsessive-compulsive disorder group. The schizophrenic group had a lower RSS average than patients with depression, patients with obsessive-compulsive disorders, or subjects in the normal control group. These results in conjunction with those of our previous studies suggest that a low RSS is specific to schizophrenia. We examined the relationship between these eye movements and neuropsychological tests and also investigated the relation between the eye movements and clinical symptoms by means of the Brief Psychiatric Rating Scale, Schedule for Affective Disorders and Schizophrenia, and the Scale for the Assessment of Negative Symptoms. The RSS correlated positively with the performance IQ and Wechsler's Maze test, but not with the Wisconsin Card Sorting Test or the verbal IQ result. The RSS also correlated negatively with negative symptoms. These results suggest that the RSS has two characteristic features: it is strongly associated with the interpersonal response and it may be connected with visuospatial and visuomotor functions including attention.

Discrimination between cardiac para- and sympathetic damage in diabetics.

To analyze the autonomic mechanism on heart rate (HR) variations, pharmacological studies were performed on 8 healthy subjects and 6 diabetic patients. Three HR variations--in supine resting position, during deep breathing in supine position (respiratory HR variations), and on standing (orthostatic tachycardia; delta HR)--were examined. The results in healthy subjects and diabetics were similar. After administration of parasympathetic blockade with atropine, respiratory HR variations were almost abolished. However, no significant difference in delta HR was found. With the addition of beta-adrenergic blockade with propranolol, delta HR was remarkably reduced. Propranolol alone did not affect the respiratory HR variations, but after propranolol administration delta HR was significantly reduced compared with that of the control. The present studies show that respiratory HR variations are predominantly mediated by parasympathicus, whereas orthostatic tachycardia is mediated by both sympathicus and parasympathicus, particularly by sumpathicus. This result suggests the possibility of discriminating the impairment of the 2 cardiac autonomic nervous systems by these simple and non-invasive tests.

The columnar and layer-specific response properties of neurons in the primary auditory cortex of Mongolian gerbils.

The columnar and layer-specific response properties of neurons in the primary auditory cortex (AI) of Mongolian gerbils were studied using single-unit recordings of responses to tone-burst stimuli presented to the ear contralateral to the recording side. During near-radial microelectrode penetrations of the AI in 100-microm steps, the best frequency (BF), best threshold (BT), best amplitude (BA), latency, tuning curve and Q10dB were recorded. Neurons encountered during single penetrations showed similar BFs, indicating a columnar frequency organization, but their latencies and Q10dBs differed. The BAs and BTs recorded within single penetrations often showed a similar value in the middle cortical layers. The latencies and Q10dBs of these neurons exhibited a tendency toward a layer-specific distribution. The latencies of neurons located in layers I-V were longer than those located in layer VI. The Q10dBs of neurons located in layers III and IV were higher than those located in layers I and VI. These results are almost consistent with those of previous studies on frequency representation, and indicated the existence of an integrative mechanism of frequency processing in the AI. This is the first study in which a layer-specific, partially columnar organization for stimulus amplitude is described.

Effects of aflastatin A, an inhibitor of aflatoxin production, on aflatoxin biosynthetic pathway and glucose metabolism in Aspergillus parasiticus.

Aflastatin A inhibits aflatoxin production by Aspergillus parasiticus via an unknown mechanism. We found that aflastatin A clearly inhibited production of norsolorinic acid, an early biosynthetic intermediate of aflatoxin, at a concentration of 0.25 microg/ml. Reverse-transcriptase polymerase chain reaction (RT-PCR), and real-time quantitative PCR (TaqMan PCR) experiments indicated that the transcription of genes encoding aflatoxin biosynthetic enzymes (pksA, ver-1, and omtA) and a gene encoding a regulatory protein for expression of the biosynthetic enzymes (aflR) were significantly reduced by the addition of aflastatin A. We also found that aflastatin A elevated the glucose consumption and ethanol accumulation by A. parasiticus, and repressed transcription of genes involved in ethanol utilization. These results suggest that aflastatin A inhibits a very early step in aflatoxin biosynthesis prior to the transcription of aflR and can influence glucose metabolism in the fungus.

Blasticidin A derivatives with highly specific inhibitory activity toward aflatoxin production in Aspergillus parasiticus.

Blasticidin A (1), an antibiotic, has strong inhibitory activity toward aflatoxin production by Aspergillus parasiticus. We prepared some derivatives of 1 and examined their biological activities. Among them, derivatives 3 and 4 without the tetramic acid moiety of 1 maintained inhibitory activity toward aflatoxin production, but did not show antifungal activity or toxicity. RT-PCR experiments indicated that derivatives 3 and 4 as well as 1 significantly reduced the expression of genes encoding aflatoxin biosynthetic enzymes (pksA, ver-1 and omtA) and a regulatory gene (aflR) in A. parasiticus. These results suggested that derivatives 3 and 4 can inhibit aflatoxin production more specifically than 1 by inhibiting an early step prior to the expression of aflR in the pathway of aflatoxin biosynthesis.

[A diffuse metastatic leptomeningeal carcinomatosis from gallbladder cancer; case report].

We encountered a 72-year-old woman with diffuse metastatic leptomeningeal carcinomatosis, who first suffered from occipital pain and died about a month after onset. On postmortem examination, gallbladder cancer (adenocarcinoma) was found to be the primary disease. We focused on its frequency and the metastatic route. On the metastatic route, we obtained the following results: tumor cells infiltrated only the cerebrospinal fluid, but not the areas surrounding the gallbladder cancer (spine or spinal cord) or into the brain parenchyma. A comparative study of the state of cerebrospinal fluid between the ventricle and the subarachnoid space disclosed that the cerebrospinal fluid pressure, cell count, and CEA and CA 19-9 levels increased more in the intraventricular cerebrospinal fluid, especially when the CEA level was higher than that in the serum. On histopathological examination, tumor emboli were seen in choroidal vessels in the ventricular wall, and tumor cells existed sparsely around choroidal secretory vessels. These results were thought to support the theory of hematogenous metastasis as Little et al proposed.

[A case of systemic lupus erythematosus associated with spontaneous bilateral epidural hematomas].

The authors reported a case of systemic lupus erythematosus (SLE) associated with bilateral epidural hematomas which had developed without any recent trauma. A 34-year-old male suddenly suffered from a severe headache and vomited several times. He had no neurological deficits on admission, but CT scans of the head revealed abnormal high density areas over the parieto-occipital regions beneath the calvarium bilaterally. At the operation, fresh epidural clots were removed. There were neither evidence of trauma nor abnormal structures which might have led to the development of the clots. He was discharged a month later, being free from any signs and symptoms. One year later, arthralgia progressed and cutaneous ulceration appeared on his feet. On the second admission, butterfly rash on the face, alopecia, polyarthritis and arthralgia, photosensitivity, systemic purpura and proteinuria were noted. With detailed immunological examinations and renal biopsy, he was diagnosed as SLE. SLE is often associated with neurologic and psychic disorders and there are some cases of intracranial hemorrhage among them. However, the association with epidural hematoma has not been reported to date. We think the degeneration of the dural vessels caused by underlying SLE resulted in the development of these epidural hematomas. We also reviewed the literature about spontaneous epidural hematoma and about bilateral epidural hematomas.